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Revisit of Imidazole–Zinc Reverse Stain for Protein Polyacrylamide Gel Electrophoresis

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Protein Electrophoresis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 869))

Abstract

Imidazole–zinc reverse stain (ZN stain) is known for its high sensitivity, ease of use, and cost-effective feature. ZN stain is compatible to many experiments of which those are proteomics-related in particular. Here, we describe the ZN staining procedures and the subsequent procedures incorporated in detail, along with the improvements of setup in aspects of visualization and documentation for post-processing ZN-stained gel images.

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References

  1. Fernandez-Patron C, Castellanos-Serra L (1990) Abstract booklet. In: Eight international conference on methods on protein sequence analysis, Kiruna, Sweden, July 1–6 1990

    Google Scholar 

  2. Fernandez-Patron C, Castellanos-Serra L, Rodriguez P (1992) Reverse staining of sodium dodecyl sulfate polyacrylamide gels by imidazole-zinc salts: sensitive detection of unmodified proteins. Biotechniques 12:564–573

    PubMed  CAS  Google Scholar 

  3. Ferreras M, Gavilanes JG, Garcia-Segura JM (1993) A permanent Zn2+ reverse staining method for the detection and quantification of proteins in polyacrylamide gels. AnalBiochem 213:206–212

    CAS  Google Scholar 

  4. Hardy E, Pupo E, Casalvilla R et al (1996) Negative staining with zinc-imidazole of gel electrophoresis-separated nucleic acids. Electrophoresis 17:1537–1541

    Article  PubMed  CAS  Google Scholar 

  5. Hardy E, Sosa AE, Pupo E et al (1996) Zinc-imidazole positive: a new method for DNA detection after electrophoresis on agarose gels not interfering with DNA biological integrity. Electrophoresis 17:26–29

    Article  PubMed  CAS  Google Scholar 

  6. Hardy E, Pupo E, Castellanos-Serra L et al (1997) Sensitive reverse staining of bacterial lipopolysaccharides on polyacrylamide gels by using zinc and imidazole salts. Anal Biochem 244:28–32

    Article  PubMed  CAS  Google Scholar 

  7. Pupo E, Lopez CM, Alonso M et al (2000) High-efficiency passive elution of bacterial lipopolysaccharides from polyacrylamide gels. Electrophoresis 21:526–530

    Article  PubMed  CAS  Google Scholar 

  8. Castellanos-Serra L, Hardy E (2001) Detection of biomolecules in electrophoresis gels with salts of imidazole and zinc II: a decade of research. Electrophoresis 22:864–873

    Article  PubMed  CAS  Google Scholar 

  9. Hardy E, Castellanos-Serra LR (2004) “Reverse-staining” of biomolecules in electrophoresis gels: analytical and micropreparative applications. Anal Biochem 328:1–13

    Article  PubMed  CAS  Google Scholar 

  10. Lin CY, Wang V, Shui HA et al (2009) A comprehensive evaluation of imidazole-zinc reverse stain for current proteomic researches. Proteomics 9:696–709

    Article  PubMed  CAS  Google Scholar 

  11. Pupo E, Lopez CM, Alonso M et al (2000) High-efficiency passive elution of bacterial lipopolysaccharides from polyacrylamide gels. Electrophoresis 21:526–530

    Article  PubMed  CAS  Google Scholar 

  12. Scheler C, Lamer S, Pan Z et al (1998) Peptide mass fingerprint sequence coverage from differently stained proteins on two-dimensional electrophoresis patterns by matrix assisted laser desorption/ionization-mass spectrometry (MALDI-MS). Electrophoresis 19:918–927

    Article  PubMed  CAS  Google Scholar 

  13. Castellanos-Serra L, Proenza W, Huerta V et al (1999) Proteome analysis of polyacrylamide gel-separated proteins visualized by reversible negative staining using imidazole-zinc salts. Electrophoresis 20:732–737

    Article  PubMed  CAS  Google Scholar 

  14. Gulin S, Pupo E, Schweda EK et al (2003) Linking mass spectrometry and slab-polyacrylamide gel electrophoresis by passive elution of lipopolysaccharides from reverse-stained gels: analysis of gel-purified lipopolysaccharides from Haemophilus influenzae strain Rd. Anal Chem 75:4918–4924

    Article  PubMed  CAS  Google Scholar 

  15. Fernandez-Patron C, Calero M, Collazo PR et al (1995) Protein reverse staining: high-efficiency microanalysis of unmodified proteins detected on electrophoresis gels. Anal Biochem 224:203–211

    Article  PubMed  CAS  Google Scholar 

  16. Fernandez-Patron C, Castellanos-Serra L, Hardy E et al (1998) Understanding the mechanism of the zinc-ion stains of biomacromolecules in electrophoresis gels: generalization of the reverse-staining technique. Electrophoresis 19:2398–2406

    Article  PubMed  CAS  Google Scholar 

  17. Lin CY, Huang HM, Chen HM (2006) Use of backlit light plate to enhance visualization of imidazole-zinc reverse stained gels. Biotechniques 41:560, 562, 564

    Google Scholar 

  18. Wu HC, Yen CC, Tsui WH et al (2010) A red line not to cross: evaluating the limitation and properness of gel image tuning procedures. Anal Biochem 396:42–50

    Article  PubMed  CAS  Google Scholar 

  19. Westermeir R, Naven T (2002) Proteomics in practice. Wiley-VCH Verlag-GmbH, Weinheim

    Book  Google Scholar 

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Acknowledgements

This work was supported by the grants from Catholic Fu-Jen University (9991A02-109931041001-2) and National Science Council of the Republic of China (NSC 99-2627-E-030-001). We thank Dr. Kuo Cheng-Yi for linguistic edition and Mr. Tsai Tzung-Lin and Dr. Wu Szu-Yu for providing gel images.

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Authors and Affiliations

  1. Catholic Fu-Jen University, New Taipei City, Taiwan

    Han-Min Chen

Authors
  1. Han-Min Chen
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Correspondence to Han-Min Chen .

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Editors and Affiliations

  1. Oklahoma Medical Research Foundation, Univ. of Oklahoma Health Sciences Center, NE. 13th Street 825, Oklahoma City, 73104, Oklahoma, USA

    Biji T. Kurien

  2. Oklahoma Medical Research Foundation, NE. 13th Street 825, Oklahoma City, 73104, Oklahoma, USA

    R. Hal Scofield

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Chen, HM. (2012). Revisit of Imidazole–Zinc Reverse Stain for Protein Polyacrylamide Gel Electrophoresis. In: Kurien, B., Scofield, R. (eds) Protein Electrophoresis. Methods in Molecular Biology, vol 869. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-821-4_43

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  • DOI: https://doi.org/10.1007/978-1-61779-821-4_43

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-820-7

  • Online ISBN: 978-1-61779-821-4

  • eBook Packages: Springer Protocols

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